1. Microbial contamination of pharmaceutical products can occur from raw materials, equipment, manufacturing environment, and personnel. Strict environmental controls and good manufacturing practices are required. 2. Sterility testing ensures pharmaceutical products intended to be sterile, such as parenteral and ophthalmic products, are free of microbial contamination. Tests include direct inoculation into culture media and membrane filtration sterility testing. 3. Validation and quality control measures are critical throughout the manufacturing process to minimize microbial contamination and ensure product sterility. These include monitoring bio burden levels, air and surface sampling, and validation of sterilization and filtration processes.

1. Lec1
Pharmaceutical
products Types
Manufacture
Microbial Contamination
Sources
Microbiological
Analysisof Air
Microbial spoilageof
pharmaceutical products
Control of microbial contamination
during manufacture
Sterile Parenteral ophthalmic otic
Non sterile Oral topical cosmetics
Raw materials Natural materials
Equipment Manufacturing
Environment
Manufacturing environment
Dry surface (wall)
Wet areas
Process operators Direct contact Skin scales
Packaging material
containers& closuresmust be bacteriologically
clean especially sterilefluids
Degradation of
active ingredients
antibioticsby B-lactamase
analgesics(Aspirin)
surfactants
Thickening & suspending agents starch
sweetening, flavoring & coloring agents sugars are labileto fermentation by acids.
preservatives
benzoic acids is decomposed byGram –ve
bacteria (Pseudomonas).
General formulation
Viscosity changes
Separation of suspended materials
Cracking of creams & emulsions
Production of toxins&pyrogens
Pyrogens & endotoxins by Gram –ve bacteria
Exotoxins by Diphtheria
Aflatoxins by toxigenic fungi
complete cycle of production of a
pharmaceutical products
includesacquisition of
raw materials
processing in to a final products
packaging
distribution.
Enviromental cleanliness&hygiene
M.O. into working surfaces & equipment
water is a source of contamination
Area should be clean & dry
Procedures for cleaning must be enforced
Good personal hygiene
Quality of starting materials water is a source of contamination
Process design product must be microbiologically acceptable
Quality control & documentation
Selection of raw materialswith low bio burden
within limit
Condition of packaging, storage & transport Should minimize or prevent contamination.
Microbial quality
The incidence& level of microbial contamination
in a pharmaceutical products
Microbial quality control
Analytical procedurescarried out on
pharmaceutical products
Hazardsof
Microbial Contamination
Spoilageof the products
health hazards to the patient
Physical
indicators
Sterilization monitoring
achieved by
In process
Contol
Heat
Sterilization
Temp record chart is compared with a master
temperaturerecord(MTR).
Gseous
Sterilization
Elevated temp is monitored for each cycle by
temp probes.
Pressure, gas concentration & humidity are
recorded
Radiation
Sterilization
A plastic dosimeter to measure radiation dose
Sterilization
by filtration
measurepore size & integrity of filter
Chemical
indicators
Brown
Tube
An ampulecontains a heat sensitivedye
changing in color
from red to greenafter time at specific temp
witness
Tube
Sealed tube contains a compound melting at the
sterilization temp
Sulphur melts at 115 ◦C.
paper strips
to determinechange in color during sterilization
cycle
Biological
indicators
standard spore suspension in water(bacillus
subtilis)
standard spores dried on a paper or plastic
carriers
placed in different sites in sterilizer.
After sterilization the spore culture is
transferred to nutrient agar
incubation and examined for developed growth
Sterility testing
for the final products
Mathematical
D or Z-value
asking for reevaluation of the sterilization
process
ensure invasion of the environment with
resistant strains.
ensures that a sterilized pharmaceutical or
medical product is free from m.o
the final control on sterilization process
not an evaluation test
passing this test increase the probability of
sterility of the products
Methods
Direct Inoculation
into nutrient media
Fluid thioglycholate
medium
for aerobic & anaerobic bacteria
Soya bean casein
digest medium
for aerobic bacteria &fungi
Membrane
Filtration
bacterial filter with pore size <0.45 um.
Sensetive
Method
for detection low level of contamination in I.V
fluids
Positive
Control
to demonstrate the growth promoting ability of
the medium
Negativetest
to ensure the sterility of the medium.
sample
size
1-2 % of batch with maximum 20 articlesselected
randomly from different parts
Steritest
a special equipment designed to permit in a closed
system the filtration of entire content of a bottle
through a pre-sterilized unit containing a 0.22 um
membranefilter. The suitable medium is added
asepticallyand unit is inoculated intact.
Advantages
a closed system to reduce false positive.
More convenient for large volume
Disadvantages
a relativelyhigh cost
Sensitive to low level contamination
Results
If no growth occurred with fresh samples, the
batch passes the test
If growth occurs, but the organism differs from
the found previously,
the test is repeated on the third Sample from the
batch
If no growth occurred with the third sample, the
batch passes the test.
not guaranteeing the sterility of the batch
an additional check , & gives confidence to the
efficacy of sterilization process.
It increasethe probability of sterility of the
product
Pyrogen Testing
Pyrogens
fever producing organicsubstance arising from
microbial contamination.
official tests for parenteral
to limit to an acceptablelevel of the risks of
infection on injecting
Rabbit test
measuring the rise in temperatureof
standardized 3 rabbits upon I.V, injection of the
sample
LAL test
LIMULUS AmebocyteLysate Test
a precise test
an extract from the blood cells of the horse Shoe
Crab (LimulusPolyphemus) contain an enzyme
coagulating in the presence of low level of
endotoxin.
only ensures that the preparation or product is
free from pyrogenic material
ensure the sterilityof the product
Advantage
Better sensitivity, specificity, reliability and
wider application
Less variation and less expensive
Aseptic area
The room should be kept under a slight positive
pressure.
Smooth walls, ceiling & floor to prevent microbial
and to erase cleaning & disinfection.
Double glass windowsDoubledoors with air
lock
Furniture & equipment should be minimal &
simple
sterile air supply and all supjects in the area
UV radiation to maintain sterility
free from infectious disease
Air must be regularly analyzed for their
microbial content.
Entry
First (Black) area Locker room non-sterile
Second(Grey) area a transient area for changing clothes
Third(White) area Aseptic area sterile gowning room
Parenteral
quality control
All parenteral products must be sterile ,so
Apply Sterility testing as a QC procedures
Explain the reasons for sterility failureeven the
article is sterilized
Sterilization control
&Sterility Assurance
Bio burden
Total number of m.o per gram or milliliter
regardlessits type
must be very low for raw materials
Water supply should be with low bio burden
prior to sterilization
Settling plates
Plates with nutrient agar are opened for a
specific time -
incubated at specific temp-coloniesare
counted.
Air sampler
Agar impingesis used , the volume of air is forced
to pass through a slit,
below which small nutrient agar is placed –
incubation-coloniesare counted
Validation of
HEPA filter
passing a certain type of oil through the filter by
a fan
then test for the presence of particulate matter
on the side.
aseptic area System suitability test
checking for the presenceof M.O.
in every site in the area